LOCUS       sequence-193973-        9199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of S. cerevisiea RPA in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9199)
  AUTHORS   Sibenaller ZA, Sorensen BR, Wold MS
  TITLE     The 32- and 14-kilodalton subunits of replication protein A are 
            responsible for species-specific interactions with single-stranded 
            DNA.
  JOURNAL   Biochemistry. 1998 Sep 8;37(36):12496-506.
  PUBMED    9730822
REFERENCE   2  (bases 1 to 9199)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Biochemistry."; date: "1998-09-8"; volume: "37(36)"; pages: 
            "12496-506"
COMMENT     Sequence Label: p11d-sctRPA
FEATURES             Location/Qualifiers
     source          1..9199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..36)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        104..208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(209..277,278..1069)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(427..446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1240..1828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1729..1748
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1982..1999
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2014..2156
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2082..2101)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2242..2264
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2258..2449)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3761..4843)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4805..4824
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4844..4921)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5127..5146
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5230..5248
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5249..5273
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             5288..5310
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        7204..7222
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7223..7247
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             7262..7284
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             8255..8277
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(8926..8944)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      8940..8987
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(9162..9167,9168..9184,9185..9190))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga
     2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg
     3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc
     3061 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg
     3121 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt
     3181 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt
     3241 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg
     3301 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat
     3361 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc
     3421 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     3481 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     3541 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     3601 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     3661 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     3721 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3781 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3841 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     3901 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     3961 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     4021 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
     4081 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     4141 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     4201 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     4261 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     4321 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     4381 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     4441 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     4501 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     4561 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     4621 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     4681 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     4741 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     4801 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     4861 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     4921 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     4981 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     5041 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     5101 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     5161 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     5221 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     5281 aaataatttt gtttaacttt aagaaggaga tataccatgg gcagtgttca actttcgagg
     5341 ggcgattttc atagcatctt caccaataag caaaggtacg ataatcccac cggtggcgtt
     5401 tatcaagttt ataacaccag gaaatctgat ggggctaaca gcaacagaaa gaatttgatc
     5461 atgatttccg atggtattta ccatatgaag gctctgttga gaaaccaagc tgcatccaag
     5521 ttccagtcaa tggaactaca aaggggtgat atcattcgcg tgataattgc agaacctgct
     5581 attgtcaggg aaagaaagaa atacgttctt ttagtagatg actttgagtt ggtccagtcg
     5641 cgtgctgata tggtcaacca aactagtact tttttggata actatttctc agagcatcca
     5701 aatgaaacct taaaagacga agatataact gacagtggta atgttgccaa tcaaacaaac
     5761 gccagcaatg ctggtgtccc tgatatgctg cattcaaact caaacttgaa tgcaaatgag
     5821 agaaaattcg ccaatgaaaa ccctaattcg caaaaaacca gaccaatttt tgccatcgaa
     5881 caactgtctc cataccaaaa cgtttggact atcaaagcaa gagtttccta caagggagaa
     5941 attaaaacgt ggcacaatca aagaggtgat ggtaaactat tcaatgtcaa cttcttggat
     6001 acctctggag aaatccgagc cacggcgttt aatgattttg ctacaaaatt taacgaaatt
     6061 ttacaagaag gcaaagtata ctatgtatca aaggcaaaac tccaaccagc taagccccaa
     6121 tttactaatc taacacaccc ttatgaactg aatttggata gagacactgt tatagaagaa
     6181 tgtttcgatg aaagtaatgt tccgaaaacc catttcaatt tcatcaaact agatgctatt
     6241 cagaaccagg aagtaaattc caacgtagac gtcctcggta ttatccaaac tataaaccca
     6301 cattttgagc taacttcaag ggctgggaag aaattcgatc gtcgtgacat cacaattgtt
     6361 gacgactctg ggttttctat ctctgttggc ctatggaatc agcaagccct tgatttcaac
     6421 cttcctgaag gttctgttgc tgccattaaa ggtgttcgtg tgacggattt tggtggcaaa
     6481 tctttgtcta tgggattttc tagtaccctg attccgaatc cagaaattcc tgaggcatat
     6541 gccttaaagg gttggtatga ttccaagggc cgcaacgcaa acttcatcac tttaaagcaa
     6601 gaacccggta tgggtggtca atcggctgct agcttaacaa aattcattgc tcagcgtatt
     6661 actattgcta gagctcaagc tgaaaatcta ggaagaagcg agaaaggtga cttttttagt
     6721 gttaaagctg ctataagttt cttaaaagtt gataattttg catatcctgc ctgttctaat
     6781 gagaattgta ataagaaagt tctggaacag cctgatggta cttggagatg tgagaagtgc
     6841 gacaccaata atgcaaggcc aaattggaga tacatcttga caatatcaat tattgacgaa
     6901 accaatcaac tatggctcac tttatttgac gaccaagcta aacaattatt gggtgttgat
     6961 gctaatacat taatgtcttt gaaggaagaa gaccccaacg aattcacaaa aattactcaa
     7021 agtatccaaa tgaacgaata tgactttagg attagagcgc gtgaggatac atacaatgat
     7081 caaagcagaa ttagatatac cgttgctaac ctacacagct tgaattacag ggctgaagcc
     7141 gactatcttg ccgatgagtt atccaaggct ttgttagctt aatggatctc gatcccgcga
     7201 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
     7261 ttttgtttaa ctttaagaag gagatataca tatggcaagt tatcaaccat ataacgaata
     7321 ttcatcagta acgggcggtg gctttgagaa ctctgagtcc cgcccaggta gtggggagtc
     7381 ggaaactaac actagagtta acaccttgac acctgtgacg atcaaacaaa ttctagagtc
     7441 caaacaggat attcaggacg gccccttcgt ttcgcataac caagaacttc atcacgtttg
     7501 ttttgtaggt gtggtgagaa acattacaga ccatactgca aatatttttt taactattga
     7561 ggatggaact ggtcaaatag aagtgagaaa atggagcgaa gatgcaaatg acttggctgc
     7621 cggtaacgat gactcttctg gtaaaggtta tggttcgcaa gtcgcccaac aatttgaaat
     7681 tggcggttac gtaaaagttt ttggtgcttt gaaagagttt ggtggtaaga aaaatataca
     7741 gtatgcggtg attaagccca tagattcatt caatgaagtg ttgacgcatc acttggaagt
     7801 catcaaatgt cattccatag ccagtggaat gatgaaacaa cctttggaga gtgcatccaa
     7861 caacaatggg caatcattat ttgtcaagga tgataacgat acatcttccg gctccagtcc
     7921 gttacaaaga attctagaat tttgtaagaa gcaatgtgag ggcaaagacg ctaattcatt
     7981 cgctgttccc attccattga tctcgcaatc cttgaatttg gatgaaacta ccgtcagaaa
     8041 ctgctgtacg accttgactg accagggttt tatctaccca acttttgatg acaataactt
     8101 ctttgcccta tgaaaatata tagattattt ttttagcatt tcttagtatt cgtttgtgta
     8161 tttctactag gtaagtacca aaatttgccg gaaatagagt ggaaaatgga accatcgaat
     8221 tcctgcagcc cgggggatcc cctctagaaa taattttgtt taactttaag aaggagatat
     8281 accatggcca gcgaaacacc aagagttgac cccacagaaa tctccaacgt caatgctcct
     8341 gtgtttagga taatagcgca aatcaaatca caacccactg aatctcagtt gattttacaa
     8401 tcgccaacca tatcatcgaa aaacggcagc gaggttgaaa tgattacgtt gaacaatatt
     8461 cgtgtatcta tgaataaaac atttgagatc gactcgtggt atgagtttgt ttgcaggaac
     8521 aatgatgacg gcgagctagg atttttgatt ctagacgctg tactatgcaa attcaaggag
     8581 aacgaagatt taagcttaaa cggtgtggtt gctttacaga gactatgtaa gaaataccca
     8641 gaaatatact agctaaaatc gcttttattc ttacctttat gtactgttgc ctctatatgt
     8701 cttcccttca tataataaat tccatttcta aaaaaattgt agttgtgttt atagatgtat
     8761 agatagataa atgaatgtgt acttgctgcc ttaacgcctt ccgctaaaaa ctttttgata
     8821 tttattttat ttttaacctt tcctgacttc aacccttact gtaatgatga tagtaggatc
     8881 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     8941 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     9001 ctatatccgg atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac
     9061 agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg
     9121 tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat
     9181 aagctgtcaa acatgagaa
//
